I/R injury may develop in conditions in which the

blood a

I/R injury may develop in conditions in which the

blood and oxygen supply to a tissue is transiently disrupted and then restored. Hepatic I/R injury is a potentially fatal complication of liver surgery (including liver transplantation). Substances that improve hypoxia tolerance selleck chemicals llc may also protect against I/R injury. CBs induce hypomotility and hypothermia, both of which result in reduced oxygen demand. The metabolic effects of CBs that promote energy storage and reduce energy expenditure (discussed previously) may also reduce oxygen demand. There is evidence that endocannabinoids acting via CB2 protect against hepatic I/R injury.99, 100 In mice, segmental ischemia followed by reperfusion (but not ischemia alone) markedly increased the hepatic levels of AEA and 2-AG, which correlated with the severity of tissue damage.99

I/R-induced tissue damage, including neutrophil infiltration and lipid peroxidation, was attenuated by pretreatment with JWH-133 in wild-type mice but not in CB2−/− mice, in which the damage was more severe than that in wild-type littermates.99 Another potent and selective CB2 agonist, HU-308, caused similar effects and also attenuated I/R-induced hepatocyte apoptosis and mitigated the tumor necrosis factor-α–induced expression of cell adhesion molecules (intercellular cell adhesion molecule 1 and vascular cell adhesion molecule 1) in hepatic sinusoidal endothelial cells.100 Thus, CB2 agonists may afford protection at multiple levels against I/R injury; this highlights their therapeutic find more potential. PD98059 CB1 receptor blockade also protects the liver from I/R injury and superimposed endotoxaemia.101 In one study, rats subjected to lipopolysaccharide plus

I/R had an immediate increase in CB1 expression in perisinusoidal hepatocytes. Rimonabant treatment reduced both tissue necrosis and serum alanine aminotransferase levels in the late phase of reperfusion and attenuated the oxidative injury.101 Further studies with peripherally restricted CB1 receptor antagonists could reinforce the therapeutic potential of this approach. Hepatic encephalopathy is a neuropsychiatric syndrome that may accompany acute liver failure. The underlying mechanisms are not completely understood, although there is evidence for the pathogenic role of ammonia, alterations in various central neurotransmitter systems, and altered cerebrovascular function. Mice with thioacetamide-induced fulminant liver failure have elevated brain 2-AG levels. The treatment of such mice with 2-AG or the CB2 agonist HU-308 improved the neurological score and cognitive function, and these effects were blocked by a CB2 antagonist. The beneficial effects of CB2 agonists could be mimicked by treatment with the CB1 antagonist rimonabant.

63 vs 375 log IU/mL, p=<0001), and more likely to be males (59

63 vs 3.75 log IU/mL, p=<0.001), and more likely to be males (59.8% vs 29.7%, p<0.001) respectively. There was no difference in HBsAg levels between those with and without hepatitis flare (3.53 vs check details 3.52 log IU/mL respectively, p=0.465). Conclusion: HBV DNA levels,

but not HBsAg levels, after HBeAg seroclearance were associated with subsequent significant viremia and hepatitic flares. Male gender and older age was associated with significant viremia. Disclosures: James Fung – Speaking and Teaching: Bristol Myers Squibb Wai-Kay Seto – Advisory Committees or Review Panels: Gilead Science; Speaking and Teaching: Gilead Science Ching-Lung Lai – Advisory Committees or Review Panels: Bristol-Myers Squibb, Gilead Sciences Inc; Consulting: Bristol-Myers Squibb, Gilead Sciences, Inc; Speaking and Teaching: Bristol-Myers Squibb, Gilead Sciences, Inc Man-Fung Yuen – Advisory Committees or Review Panels: GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer; Grant/Research Support: Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science The following people

have nothing to disclose: Danny Wong Background Selleckchem Deforolimus and aims: Chronic hepatitis B (CHB) patients with moderate to severe liver disease need to be treated. Currently, little is known about the association between the severity of the liver disease and both host factors (including IL28B rs12979860) and viral factors such as mutations in Basal Core Promoter (BCP) and Precore (PC) regions, HBsAg and HBV-DNA levels and HBV genotypes. Therefore, selleck we investigated these relationships in a large cohort of unselected, well-characterized, treatment-naïve CHB patients.

Methods: 406 HBsAg positive patients were included. Liver biopsy was available for all patients. HBsAg and HBV-DNA levels, HBV genotypes, BCP and PC variants were determined the day of the liver biopsy. BCP and PC variants were detected by reverse hybridization using the Inno-LIPA HBV PreCore assay (Innogenetics). Histo-logical lesions were assessed using the METAVIR scoring system. Results: Out of the 406 CHB patients: 101 were HBeAg(+), 305 HBeAg(-). Fibrosis stage F0-1, F2, F3 and F4 was observed in 61% 23%, 8% and 8% of the patients, respectively. PC, BCP and BCP+PC variants were found in 25%, 29% and 28% of the patients, respectively. The HBV genotype distribution was: A, 26%; B, 11%; C, 9%; D, 24% and E, 30%. The IL28B genotype distribution was: CC, 43%; CT, 31% and TT, 26%. Patients with fibrosis stage ≥F2 had higher ALT level (3.28±4.93 times of the normal) as compared to patients with F0-1 fibrosis (1.70±2.07) (p<0.

healthy

controls 2) clinical factors associated with EAT

healthy

controls 2) clinical factors associated with EAT and the association with the severity of NAFLD 3) whether EAT predicts early atherosclerotic vascular damage, evaluated by common carotid arteries intima-media thickness (CC-IMT), and subclinical cardiac dysfunction. Methods: EAT thickness, i. e. echofree space between the outer wall of the myocardium and the visceral layer of pericardium, was evaluated by transthoracic echocardiogram in 155 consecutive patients, 43 with biopsy proven NAFLD (mean age 51 ±11 years), PF-562271 mw and in 87 healthy controls (mean age 52± 11 years), in whom hepatic steatosis was excluded by abdominal ultrasonography. In all patients complete anthropometric, clinical and biochemical data were obtained, and CC-IMT evaluated. Results: Patients with NAFLD had higher EAT thickness than controls (5.1 ±2.6 vs.3.0±2.0 mm, p=0.001). At univariate analysis, BMI (OR 1.16, 95% CI1.33), fasting glucose >100 (OR 4.2, 95% CI 1.14-20), HbA1c (OR 2.6, 95%CI 1.19-8.3), diabetes (OR 1.28, 95% CI1.73), and metabolic syndrome (OR 1.34, 95% CI 1.151.57) were significantly PF-6463922 nmr associated with increased EAT thickness (above median value). In the 43 patients with liver histology (15 simple steatosis, 25 nonalcoholic steatohepatitis without cirrhosis, and 3 NAFLD with cirrhosis), EAT thickness increased with the severity of steatosis (p=0.01). Increased

EAT thickeness was associated with CC-IMT >0.65 mm (median value of controls: OR 4.6, 95% CI 1.6-15.9). Furthermore, EAT thickness was inversely correlated with cardiac early diastolic dysfunction, as detected by the early/atrial peak flow ratio (E/A ratio) (OR 0.07, 95% CI 0.01-0.38). Conclusions: EAT thickness

is higher in NAFLD patients than in healthy controls, is associated with adiposity and insulin resistance, and with early markers of cardiovascular risk. Further studies in large cohorts are required to define whether EAT thickness represents an easily assessable diagnostic tool to predict cardiovascular risk. Disclosures: The following people have nothing to disclose: Anna Ludovica Fracanzani, Giuseppina Pisano, Rosa Lombardi, Luca Valenti, Cristina Bertelli, Tiziana Tonella, Ferdinando Massari, Andrea Baragetti, Liliana Grigore, Alberico selleck Catapano, Silvia Fargion Introduction: Insulin resistance and inflammation are hallmarks of NASH. In addition to the well-known effects on carbohydrate metabolism, insulin resistance and inflammatory cytokines have profound effects on protein and amino acid metabolism. The aim of this study was to develop detailed amino acid metabolism signatures across the histologic stages of NAFLD. Methods: Five groups of participants were studied: 1) lean controls (n=20), 2) obese with normal histology (n=10), 3) obese with simple steatosis (n=10), 4) obese with NASH (FS 0-3)(n=20). Obese patients were matched for BMI, age and gender. Al participants were matched for age and gender.

0 mg daily) Duration of treatment in study ETV-901 was at the di

0 mg daily). Duration of treatment in study ETV-901 was at the discretion of the investigator; patients could continue until study closure if the investigator judged that continued treatment was in the patient’s best interest or discontinue at any time. All patients who discontinued treatment in ETV-901 selleckchem were required to be followed for at least 24 weeks postdosing to assess safety. The studies were conducted in accordance with the ethical principles of the Declaration of Helsinki and in adherence to the laws and regulatory requirements of all participating countries.

Written informed consent was obtained from all study participants. Complete inclusion criteria for enrollment in the ETV-022 study have been described.18 Eligible patients were 16 years or older and had HBeAg-positive CHB and compensated liver function. Patients were required to have detectable hepatitis B surface antigen (HBsAg) for at least 6 months and evidence of chronic hepatitis on a baseline liver biopsy completed within 1 year of randomization. At screening, patients were required to have serum HBV DNA ≥3 MEq/mL (≈3 million copies/mL) by bDNA assay, and alanine aminotransferase

(ALT) 1.3-10 times the upper limit of normal (ULN). Exclusion criteria included coinfection with hepatitis C virus, hepatitis D virus, or human immunodeficiency virus (HIV), prior treatment with lamivudine for >12 weeks, and exposure to INCB024360 datasheet other antiviral agents within 6 months of randomization. Patients treated in study ETV-022 who were eligible to enter study ETV-901 are described above in Study Design. For the HBeAg-positive entecavir long-term cohort, time on treatment for efficacy analyses was defined as the total duration in weeks from the first dose in ETV-022 to the last date of dosing up to Week 240 (Year 5) in ETV-901. Safety analyses were based on exposure during ETV-901. In ETV-901, serum HBV DNA was determined by polymerase chain reaction (PCR) assay at 12-week intervals during the first year and at 24-week intervals thereafter while treatment

continued. HBV serologies were obtained every 12 weeks during the first and second find more year of open-label treatment. Efficacy assessments for the cohort include proportions of patients at 48, 96, 144, 192, and 240 weeks (Years 1, 2, 3, 4, and 5, respectively) who met the following endpoints: HBV DNA <300 copies/mL, HBeAg loss, HBeAg seroconversion, and normal ALT (≤1.0 × ULN). Mean serum levels of HBV DNA and ALT for the cohort were also determined at baseline and at 24-week intervals through Year 5. Safety analyses for the HBeAg-positive entecavir long-term cohort included the following events occurring on-treatment during study ETV-901: adverse events, serious adverse events, treatment discontinuations due to adverse events or laboratory abnormalities, and ALT flares. Deaths that occurred on-treatment in study ETV-901 or during off-treatment follow-up are also described. The results of safety analyses for study ETV-022 have been reported.

HSC share many features with professional antigen-presenting cell

HSC share many features with professional antigen-presenting cells (APCs),11 and have been shown to process and present antigen to T cells.12 T cell

activation requires Caspase inhibitor clinical trial interaction of the T cell receptor (TCR) with cognate peptide antigen presented in the context of major histocompatibility complex (MHC) on APC, as well as the interaction of ligand-receptor pairs providing costimulatory signals. Once T cells are activated, coinhibitory molecules play a role in dampening the T cell response, serving as an “off-switch.” APC regulate T cell responses through the balance of signals delivered through costimulatory and coinhibitory molecules, with B7 family ligands expressed on APC playing a major role in T cell mediated immunity.13 Accumulated evidence demonstrates that there are at least seven members present in the B7 family, namely, B7-1

(CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2, B7-H3, and B7-H4.14 B7-1 and B7-2 interact with the receptor CD28 on T cells to provide costimulatory signals as well as interacting with CTLA4 to provide coinhibitory signals. B7-H1 LDK378 price and B7-DC interact with PD-1 to induce T cell apoptosis. B7-H2 interacts with ICOS on T cells and provides a costimulatory signal. B7-H3 and B7-H4 are newly identified ligands that inhibit T cell responses by interacting with as yet unidentified receptors.15 In the present study we investigated the impact of AHSC on adaptive

immune responses in the context of B7 family members using an in vitro mouse model. AHSC express the selleck coinhibitory molecule B7-H4, which provides a signal to dampen antigen-specific T cell responses. This work bears important implications for the dysfunctional immune responses that are often described in liver fibrosis, which is the natural environment of AHSC, and suggests a major role of these cells in modulating T cell immunity. α-SMA, alpha smooth muscle actin; AHSC, activated hepatic stellate cells; APCs, antigen presenting cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sort; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GFAP, glial fibrillary acidic protein; HCV, hepatitis C virus; HSC, hepatic stellate cells; IFNγ, interferon gamma; IL-2, interleukin 2; LCMV, lymphocytic choreomeningitis virus; MAPK, mitogen activated protein kinase; MFI, mean fluorescence intensity; MHC, major histocompatibility complex; QHSC, quiescent hepatic stellate cells; TCR, T cell receptor. C57BL/6 mice (Jackson Laboratory) were used for HSC isolation. CD8+ T cells specific for lymphocytic choreomeningitis virus (LCMV) glycoprotein gp33-41 were isolated from spleens of P14 TCR transgenic mice. HSCs were isolated from mouse livers as described.

38% 2-year recurrence- free survival, p=0 0003) Principal Compo

38% 2-year recurrence- free survival, p=0. 0003). Principal Components Analysis discriminated cirrhotic and HCC see more tissues, and HCC patients

with poor (<2 year) vs. good (>2 year) recurrence-free survival. Loss of CDH1 expression correlated with up-regulation of hepatocyte proliferation promoters MET and YAP1. CDH1, MET, and YAP1 were independent predictors of recurrence-free survival by Cox regression when corrected for tumor stage (p<0. 0001). Conclusion: HCV-cirrhosis is characterized by proliferation of liver stem cells and inhibition of hepatocyte proliferation. HCC tumors in which this pattern persists have superior outcomes to those which acquire a hepatocyte proliferation signature (loss of CDH1 and MST1, gain of MET, YAP1, MCM2). Genes in this signature should be studied further for potential as tissue or serum biomarkers for patient risk stratification. CDH1 and MET are candidates for personalized therapies with targeted pharmaceutical agents. Cox proportional hazards modeling of expression levels of proliferation genes, corrected for stage at diagnosis. The final model was highly significant (p<0. 0001) Parameter Parameter Tyrosine Kinase Inhibitor Library manufacturer Estimate Chi-Square p-value Hazard Ratio Stage at diagnosis 1. 5 25. 5 <0. 0001 4. 48 CDH1 −1. 09 6. 75 0. 009 0. 34 MET 1. 11 3. 23 0. 07 3. 05 YAP1 1. 61 5. 74 0. 017

5. 01 Disclosures: The following people have nothing to disclose: Martha K. Behnke, Mark Reimers, Robert A. Fisher INTRODUCTION: Intrahepatic Cholangiocarcinoma (ICC) is a rare bile duct cancer with dismal prognosis. Fusion events are among the most potent oncogenic drivers, this website and recent studies report dramatic therapeutic responses blocking these targets in melanoma and lung cancer. We aimed to identify fusion events that meaningfully contribute to ICC pathogenesis. METHODS: We analyzed a cohort of 115 ICC cases: 7 ICC

fresh-frozen paired samples were screened for fusion events using RNA-seq (HiSeq2000sequencer), and 108 paraffin-embedded tissues were used to validate the finding by RT-PCR and sanger sequencing. To identify fusion events, raw cDNA reads were aligned to a reference genome, with subsequent filters applied via in-house methods. A fusion gene was selected based on the number of supporting reads and partner genes and validated in the same patient where it was identified. Whole-genome sequencing was run in the sample with the fusion event. NIH3T3 cells were stably transfected to over-express the full fusion gene. The effect of the fusion gene on cell migration was investigated in vitro (transwell assay). RESULTS: An interchromosomal event resulting in the formation of a fusion gene comprising portions of an oncogenic tyrosine kinase receptor, FGFR2 (10p12) and a gene involved in epithelial differentiation, PPHLN1 (12q12) was identified in 1 patient.

The observed effects of leaf wetness and temperature on infection

The observed effects of leaf wetness and temperature on infection by P. melanocephala could help explain the initiation, rate of increase and decline of brown

rust epidemics in the field. “
“Detached Vitis vinifera (cv Chardonnay) inflorescences were inoculated with spore suspensions of either Colletotrichum acutatum or Greeneria uvicola at 25°C, and a combination of light microscopy, scanning electron microscopy and plating out of inoculated flowers on to dichloran rose bengal chloramphenicol agar was used to investigate the time frame for infection. Colletotrichum Gefitinib acutatum infection commenced within 2 h of inoculation, while infection by G. uvicola commenced between 12 and 18 h postinoculation. All parts of the flowers were infected by

both fungi. “
“Quarantined phytopathogens such as Pepino mosaic virus (PepMV) and Clavibacter michiganensis subsp. michiganensis can be introduced into tomato transplant houses and fields on infested seed and thereby cause significant economic losses. Hence, specific and sensitive seed health assays are required to exclude these organisms. Currently, separate assays must be conducted on seed samples for each pathogen, making the process time-consuming and expensive. One approach to improve the efficiency of seed health testing is multiplex real-time PCR; however, PCR can be inhibited by compounds co-extracted from seed tissues with nucleic acids. To address this concern, we explored the use of magnetic capture hybridization (MCH) to PD98059 cell line concentrate and purify target nucleic acids prior to real-time PCR. The combination of MCH with multiplex real-time PCR resulted in a 102–103-fold increase in detection sensitivity for both pathogens compared to DNA extraction and direct selleck screening library multiplex real-time PCR. The detection threshold observed for the MCH multiplex real-time PCR assay was a combination of 105C. michiganensis subsp. michiganensis CFU/ml plus a 10−4-fold dilution of total RNA extracted from PepMV-infected tomato leaf tissue. These observations provide proof for the concept that MCH can facilitate the simultaneous detection of Clavibacter michiganensis subsp. michiganensis and

PepMV by multiplex real-time PCR. “
“This study examined the effect of ASD strain (Aspergillus flavipes), isolated from continuous cropping soil for pepper and named by the sampling position, on soil microflora and soil enzymes in rooting zone soil of healthy and diseased (Phytophthora capsici) pepper plants. Results showed that the ASD strain could significantly reduce the number of bacteria and actinomycetes, with a significant increase in fungi in the rhizosphere soil of both healthy and diseased plants. With increasing colonization time of the ASD strain, the number of bacteria and actinomycetes decreased initially and then increased gradually, while the number of fungi was first increased significantly and later decreased slowly.

Conclusions:  Wire

assisted transpancreatic septotomy is

Conclusions:  Wire

assisted transpancreatic septotomy is a safe and effective alternative technique to traditional NK in patients who have failed standard BC techniques. It also allows other pre-cut techniques such as NK to be used should initial WTS be unsuccessful. “
“The prelims comprise: Half-Title Page Companion Website Title Page Copyright Page Table of Contents List of Contributors Preface “
“Damage to the hepatic vessels can be caused by iatrogenic factors, systemic diseases, prothrombotic disorders or hereditary hemorrhagic telangiectasia. Several uncommon clinical entities are related to these vascular disorders. (a) ischemic cholangiopathy, due to impaired hepatic arterial inflow, consists of necroses and/or stenoses of the bile ducts, or ductopenia; (b) Budd–Chiari syndrome (mostly due to thrombosis of the hepatic Vemurafenib order veins or terminal portion of inferior vena cava); and (c) sinusoidal obstruction syndrome – venoocclusive disease (mostly due to toxic injury), both cause ascites, portal hypertension and marked hepatic dysfunction. (d) Acute portal vein thrombosis causes acute abdominal pain and may be complicated by intestinal infarction. (e) Noncirrhotic portal hypertension can be caused by chronic extrahepatic portal vein obstruction (generally due

to thrombosis), obstruction of hepatic microcirculation (including schistosomiasis), or arterioportal fistula. “
“Aim:  Mast cells may be involved in the pathogenesis of nonalcoholic selleck chemical steatohepatitis (NASH). The mast cell protease click here chymase contributes to the formation of angiotensin II and matrix metalloproteinase (MMP)-9, both of which are intimately involved in liver fibrosis. Therefore, we hypothesized that chymase plays an important role in the development

of NASH. Methods:  Hamsters were fed a methionine- and choline-deficient (MCD) diet for 8 weeks. These animals were divided into two groups and received either TY-51469 (1 mg/kg per day) or placebo. A third group was fed a normal diet as a control. Results:  Total plasma bilirubin, triglycerides, and hyaluronic acid levels were significantly higher in the MCD diet-fed hamsters than in the normal diet-fed hamsters, but the levels were significantly lower in chymase inhibitor-treated MCD diet-fed hamsters than in placebo-treated MCD diet-fed hamsters. Using histological analysis, marked steatosis and fibrosis were observed in MCD diet-fed hamsters, but these changes were significantly attenuated by treatment with the chymase inhibitor. Increases in mast cells and chymase-positive cells were observed in the liver after the MCD diet, but the increases disappeared in the chymase inhibitor-treated group. The significant increase observed in chymase activity in liver tissue extract from the MCD diet-fed group was also reduced by treatment with the chymase inhibitor. Chymase inhibition significantly reduced not only angiotensin II expression but also matrix metallopeptidase 9 activity in MCD diet-fed hamsters.

The first few subjects who consented

did so in the full k

The first few subjects who consented

did so in the full knowledge that the dose they would receive was not likely to give selleckchem them any benefit and that they would not be able to have an additional dose of the same vector as their immune system would then reject any subsequent vector particles. Nevertheless, two of our patients, who were motivated purely by altruistic desire to help the progress of treatment for their condition, volunteered and so received the low-dose treatment. The first subject had no adverse reaction acutely to the infusion and his factor IX level stabilized at 2%. Consequently, for the last 2 years since the vector infusion, he has been able to stop prophylaxis and only treats himself for accidental injuries (Fig. 1). The second subject, treated with the low dose, 2 × 1011

vector genomes/kilogram body weight (vg/kg), also achieved a stable baseline of 2%, but due to his much more Birinapant mw severe pre-existing joint damage has needed to continue on prophylaxis, albeit with decreased frequency of dosing (Fig. 2). As both subjects had attained levels of less than 3% the next two subjects were treated at the intermediate dose level of 6 × 1011 vg/kg. Subject 3 attained a stable base line of 2% but also due to pre-existing joint damage has needed to continue prophylaxis but at greatly increased intervals of up to 3 weeks, compared to twice weekly before gene therapy. Subject 4 attained a base line level of 3%, which has persisted for 15 months such that he has needed no factor infusion for a year. Earlier treatments were for accidental injury provoked bleeding (Fig. 3). As neither of the subjects had achieved a base line level above 3% the next two subjects were treated at the highest dose level of 2 × 1012 vg/kg. Subject 5’s course is shown in Fig. 4. Having stabilized in the factor IX range of 5–7% a sharp rise in the liver marker enzyme, ALT, to over 10 times baseline occurred, concomitant with a fall of

the factor IX level to 3%. A course of Prednisolone starting at 60 mg/day and rapidly tapering over 6 weeks was given. ALT fell rapidly to baseline and factor IX levels rose to 6%. However, over the following months his baseline level has fallen to stabilize at 2%. The subject has not needed prophylaxis for see more 15 months since gene vector infusion but has occasional trauma provoked bleeding for which he treats himself with replacement therapy on demand. Because the rise in transaminase level occurred in subject 5 after 6 weeks the next subject had already been treated at the same dose level. His course is shown in Fig. 5. Following vector infusion the subject’s factor IX level rose to 8–10%. At 8 weeks, a slight rise in liver enzymes was noted and when the factor IX level fell to 3% he was started on a short course of Prednisolone.

The first few subjects who consented

did so in the full k

The first few subjects who consented

did so in the full knowledge that the dose they would receive was not likely to give buy GDC-0973 them any benefit and that they would not be able to have an additional dose of the same vector as their immune system would then reject any subsequent vector particles. Nevertheless, two of our patients, who were motivated purely by altruistic desire to help the progress of treatment for their condition, volunteered and so received the low-dose treatment. The first subject had no adverse reaction acutely to the infusion and his factor IX level stabilized at 2%. Consequently, for the last 2 years since the vector infusion, he has been able to stop prophylaxis and only treats himself for accidental injuries (Fig. 1). The second subject, treated with the low dose, 2 × 1011

vector genomes/kilogram body weight (vg/kg), also achieved a stable baseline of 2%, but due to his much more BTK inhibition severe pre-existing joint damage has needed to continue on prophylaxis, albeit with decreased frequency of dosing (Fig. 2). As both subjects had attained levels of less than 3% the next two subjects were treated at the intermediate dose level of 6 × 1011 vg/kg. Subject 3 attained a stable base line of 2% but also due to pre-existing joint damage has needed to continue prophylaxis but at greatly increased intervals of up to 3 weeks, compared to twice weekly before gene therapy. Subject 4 attained a base line level of 3%, which has persisted for 15 months such that he has needed no factor infusion for a year. Earlier treatments were for accidental injury provoked bleeding (Fig. 3). As neither of the subjects had achieved a base line level above 3% the next two subjects were treated at the highest dose level of 2 × 1012 vg/kg. Subject 5’s course is shown in Fig. 4. Having stabilized in the factor IX range of 5–7% a sharp rise in the liver marker enzyme, ALT, to over 10 times baseline occurred, concomitant with a fall of

the factor IX level to 3%. A course of Prednisolone starting at 60 mg/day and rapidly tapering over 6 weeks was given. ALT fell rapidly to baseline and factor IX levels rose to 6%. However, over the following months his baseline level has fallen to stabilize at 2%. The subject has not needed prophylaxis for see more 15 months since gene vector infusion but has occasional trauma provoked bleeding for which he treats himself with replacement therapy on demand. Because the rise in transaminase level occurred in subject 5 after 6 weeks the next subject had already been treated at the same dose level. His course is shown in Fig. 5. Following vector infusion the subject’s factor IX level rose to 8–10%. At 8 weeks, a slight rise in liver enzymes was noted and when the factor IX level fell to 3% he was started on a short course of Prednisolone.